A versatile EPR toolbox for the simultaneous measurement of oxygen consumption and superoxide production.

ABSTRACT: In this paper, we describe an assay to analyze simultaneously the oxygen consumption rate (OCR) and superoxide production in a biological system. The analytical set-up uses electron paramagnetic resonance (EPR) spectroscopy with two different isotopically-labelled sensors: ¹⁵N-PDT (4-oxo-2,2,6,6-tetramethylpiperidine-d₁₆-¹⁵N-1-oxyl) as oxygen-sensing probe and ¹⁴N-CMH (1-hydroxy-3-methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine, a cyclic hydroxylamine, as sensor of reactive oxygen species (ROS). The superoxide contribution to CMH oxidation is assessed using SOD or PEGSOD as controls. Because the EPR spectra are not superimposable, the variation of EPR linewidth of ¹⁵N-PDT (linked to OCR) and the formation of the nitroxide from ^14</

SUBMITTER: Donatienne d'Hose 

PROVIDER: S-EPMC7804984 | biostudies-literature | 2021 Apr

REPOSITORIES: biostudies-literature